Method of stimulating the growth of mammary glands and lobulealveolar system of mammals



Patented Mar. 18, 1952 UNITED STATES ATENT OFFICE Charles W. Turner,Columbia, Mo-., assignor to American Dairies, Incorporated; Kansas City,

a corporation of Maryland, and The Quaker Oats Company, Chicago, .IiL, acorpo ration of New Jersey No Drawing. Application r lipril 3, 1948,Serial No. 18,857

11 Claims.

This invention relates to improvements in stimulating. the growth ofmammary glands and the initiation of milk secretion in livestock by theaddition to their ration of selected natural or synthetic chemicals andhormones or derivatives of the same.

In brief, the invention contemplates the following basic concepts:

-1. The use of progesterone or related compounds havingprogesterone-like activity alone orin admixture to stimulate the growthof the lobule-alveolar system of the udder.

2. The use of progesterone or related compounds having progesterone-likeactivity with estrogens to stimulate the growth of the lobulealveolarsystem of the udder.

3. The use of progesterone and estrogen with protamone to stimulate thegrowth of the milk secreting system of the udder.

4. The use of. progesterone and estrogen with androgen to stimulatethe-same type of growth.

5. The use of progesterone, estrogen, androgen and protamone to producemaximum lobule-alveolar growth.

These compounds may be used on normal nonpregnant animals,sterile-females and some males as well as pregnant animals with poorgenetic udder size potential. I

A modification of the invention resides in the concept of inducing thestate of lactation-after the above growth stage has been fullydeveloped.This isaccomplished as follows:

i 1 By discontinuing the administration of progesterone or compoundshaving progesterone-like activity.

2. Increasing the amountof estrogen or estrogen-like compounds whichwillinitiate lactation and regular milking will be inaugurated, then stop orat least reduce the amount of estrogen.

3. A procedure as in paragraph 2 above, with the; addition of aprotamone to the estrogen: to bring inf-ull lactation.

4. A procedure as in paragraph 2 abovewhereina male hormone is added asa supplemental factor.

5. The administrationof estrogen, androgen and protamone to initiatelactation, then reducingthe" estrogen and androgen while increasing theprot'amone to full lactation according to the body weight. In otherwords theinvention resides primarily iniincorporating, into the rationofmammalsone or: a. series. of hormone or hormone-like: compounds which;havebeen found to synergize with eachzother to the-end thatm'aximum andcom-- plete mammary gland growth is effected and-lam tation isstimulated or improved. The amounts of the hormone prescribed, as wellas the ratioof one to the other, are critical if maximum growth is to beattained. The uniform oral'administration of thehormones andhormone-like compounds as an integral part of the ration furnishesuniform amounts of the hormones in the blood and tissues comparable tothe endo'g enou's supply of the essential factors. In addi-- tion tothe' administrati'on of the hormones and other substances suggested, thesuccess of the in vention depends as well upon the proper timing orsequence of administration of these substances or compositions.

In its broader aspects the invention contem plates the improvement inmilk producing capacity of normal animals whose mammary'gland' growthduring pregancy is deficient in the natural endogenous hormones whichnormally stimulat'e the growth of these glands. It furtherco'ntemplatesthe stimulation and growth of mainmary glands of sterile animals orthose incapable of a normal pregnancy during which time complete growthof the mammary gland normally occurs.

In animals which abort or lose the products of conception before the endof the normalpe+ riod of pregnancy, this invention provides a method ofimproving the growth and capacity of the-udder and its milk producingsystem. Bythe' use of the substances suggested and hereinafter describedin more detail, the tendency of such animals to abort-may be preventedor delayed.

This invention is an improvement upon an application Serial No. 587,444,filed April 9, 1945, issued February 13, 1951, as Patent No. 2,541,447,and a continuation-in-part of an application Serial No. 636,954, filedJuly 29, 1946, now abandoned.

It is also contemplated that with the hormones and other compoundssuggested there may be admixed salt, mineral mixtures of calcium,phosphorous,-anad trace elementssuch asfeedingoils of various types orother supplements. Where these additional materials are added the-totalsupplement either separately or mixed with the feed concentrate shouldbe such as to provide the administration ofthe hormones andchemicals'within the range-of daily intake specified in exampleshereinafter provided, as well as according to the teaching with respectto the proper time, sequence for administering the different substancesor compositions.

the like.

3 GROWTH OF THE MAm/IARY GLAND AND MILK SECRETION It is well known tothe art that in normal animals the growth of the mammary glands (themilk secreting glands) occurs in two stages:

(a) When animals become sexually mature, certain endogenous hormonesgradually stimulate the growth of the duct system which is in generalappearance, comparable to the naked branches of a tree. Glands at thisstage are capable of being stimulated so as to produce a slight milksecretion.

(b) When a mammal conceives, the endogenous hormones secreted duringpregnancy stimulate further extension of the duct system and in additionthe growth of enlarged end buds called the lobule-alveolar system. Thislatter type of growth of the gland vastly increases the number ofepithelial cells which secrete milk and thus greatly increases thecapacity of the mammary glands to secrete milk.

Shortly before normal parturition the growth phase of the mammary glandnow beingcomplete, the endogenous hormones appear to selectively actupon and stimulate the epithelial cells to gradually increase milksecretion which is further greatly increased after parturition.Unfortunately, there are great differences in the amount-of endogenoushormones which stimulate the growth and milk secretions of the mammaryglands in various strains, breeds and classes of animals. In some stockthe mammary glands (udder) are very small in size and never attainoptimum functional condition, whereas in other animals the extent ofgrowth of secreting tissue is very great. Due in part to the variationin the amount of gland tissue, there is a variation in the yield ofmilk.

This invention has to do with the method of increasing the extent ofgrowth of the mammary glands by supplementing the normal endogenoushormones of animals whose glands are deficient in this respect. Manydairy animals each year fail-to conceive. In others when the lactationprocess stops (go dry) they become worthless as dairy animals dueto ourpast inability to, reinitiate growth of the mammary gland comparable tothat occurring during pregnancy and lactation comparable to thatobserved after parturition. By the use of the substances suggested andutilizing such substances according to the teachings hereinafter ,setforth, such animals may be again brought into intense milk secre- .tion;

' PROGESTERONE AND RELATED q r v COMPOUNDS standard in the biologicalassays of these chemical compounds; 1 mg. of progesterone equals oneinternational unit. A number of other related compounds haveprogesterone-like activity, in-

cluding pregneninolone, desoxycorticosterone, dehydroandrosterone,acetoxy-pregneninolene and These compounds and others possess varyingdegrees of progesterone-like activity and may be substituted forprogesterone in the various examples according to the amount of oralprogesterone-like activity which they possess.

4 NATURAL AND SYNTHETIC ESTROGENIC OR FEMALE SEX HORMONES Estrogenichormones are characterized by their specific influence upon the genitaltissues of the female, includin growth of the vagina, stimulation of theuterus, growth and secretion of the oviduct in birds. The most potentnatural estrogens include estradiol, estrone, estriol, equilin,hippulin, equilenin, and their derivatives.

The most potent estrogen-like chemical compounds include diethylstilbestrol and its derivatives such as its esters and ethers; dihydrostilbestrol (hexestrol), as well as derivatives of the latter;benzestrol, 2-4 di(parahydro-xyphenyl 3- ethylhexane) and itsderivatives. These compounds are mentioned in copending applicationSerial No. 587,444 and vary in oral physiological activity and differ inpotency from one another. Their use will be described in physiologicalterms using dimethyl ether of diethyl stilbestrol as a standard. Othercompounds of a similar nature vary in effectiveness, depending upontheir oral potency.

NATURAL AND SYNTHETIC THYRO-ID HORMONE OR THYROXINE The thyroid glandsecretes a hormone called thyroglobulin which contains the amino acidcalled thyroxine as the active physiological ingredient. The entiredried thyroid gland can be administered or the thyroglobulin can beseparated and purified. In any event, thyroxine or its derivatives servethe same physiological purpose.

Thyroxine can be formed in vitro in any pro tein containing tyrosine bya process described in U. S. Patents 2,329,445 and 2,379,842. Thematerial formed, according to the teachings of these patents, is, forconvenience in the art, designated as a protamone and contains about 3%of thyroxine which will be used as a standard in the examples, but equalsynergistic effect may be obtained by the oral administration of otherthyro-active preparations in suitable proportions.

TESTOSTERONE, ANDROSTERONE, AND RELATED lWALE HORMONES Androgenichormones are characterized by their specific influences upon thesecondary sex glands of male mammals and upon the growth of the comb andhead furnishings of both male and female fowls. The most potentcompounds having these general properties include testosterone and itsderivatives, androsterone and its derivatives and many related compoundshaving varying physiological effects. These hormones are normallysecreted by both male and female mammals and birds and are excreted invarious forms in the urine and feces.

As described in copending application Serial No. 686,954 filed July 29,1946, the presence of large quantities of androgens has been observed inthe feces of lactating dairy cattle. Dried manure from lactating cows,crude extracts of cow manure and purified and separated androgenichormones may be substituted on an equivalent oral hormone basis in theexamples.

COIWPLETE GROWTH OF THE MAME/TARY GLAND OF NORMAL, STERILE OROVARIECTOMIZED MAMMALS The growth of the female (and some male) mammaryglands is stimulated to extensive duct development and somelobule-alveolar growth by feeding the regular grain mixture to which isadded specified amounts 'of any one of the "hill estrogens such asdiethyl stilbestrol or its derivatives, as described in Serial No.587,444. The growth of the mammary gland of a similar type and ofvariable extent can be stimulated by the inclusion in the feedcomposition of androgens as described in Serial No. 586,954.

' These hormones either alone or together still fail to complete thegrowth of the lobule-alveolar system of the mammary gland comparable tothat observed after the middle of prenancy. This important stage of themammary gland development can be stimulated by the incorporation in thefeed composition of either progesterone, pregneninolone or other naturalor synthetic compounds having the property of producing progestationalproliferation of the rabbit uterus. For the most efficient action of theprogesterone or the other compounds specified, small amounts of thenatural or synthetic estrogens should be added to the feed composition.Thus, while progesterone when fed alone in suitable amounts willstimulate complete lobule-alveolar growth, it has been discovered thatabout 20% as much progesterone is required when estrogen in minimalamounts is administered simultaneously and acts synergistically upon themammary gland.

COMPOSITIONS OF MATTER AND TIME SE- QUENCE OF THEIR ADMINISTRATION (a)Normal,

(1)) Non-pregnant, (0) Pregnant,

(d) Sterile.

According to the invention set forth in Serial No. 587,444, feedcompositions containing natural and synthetic estrogens were describedhaving the function of stimulating the growth of the mammary glandsystem and varying amounts of the lobul'e-alveolar system. The extent ofthis development compares with the development of' the udders of heifersand of cows which have been dried for a number of months.

h It has been determined that it is possible to stimulate the growth ofmammary gland development and maintain the growth of the lobulealveolarsystem comparable to the extent of development which occurs in the udderduring nor- .mal pregnancy, or to stimulate an increased or augmentedtype of growth in the glands of pregnant animals if the endogenoushormone system is'deficient by the synergism of several hormones,including estrogen and progesterone.

GROWTH OF THE UDDER The growth of the udder may be stimulated in variousWays, as follows:

1. The duct. system of the udder (or mammary gland) may first bestimulated to extensive duct growth by means of substances containinge'strogens, such as dimethyl ether of. diethyl stilbestrol or ofandrogens of various typesand derivatives of the same in varying amountsof estrogens and androgens administered simultaneously as an integralpart of the feed or ration.

6 2. The duct and lobule-alveolar "system" of the udder may be developedcontinuously by the'administration of progesterone or progesterone-likecompounds alone, or with estrogen or synthetic estrogen-like compoundssimultaneously administered without first developing the duct system. 3.During normal pregnancy, the growth ofthe lo'bule-alveolar system isstimulated by endogenous hormones. The extension of the udder or mammarygland ma beeffectedin cases of deficient endogenous hormone secretion,byoral administration of compositions of matter containing progesteroneor progesterone-like "compounds and estrogen or estrogen-like compoundssimultaneously.

INITIATION OF MILK SECRETION PARTURITION Innonpregnant animals thegrowth of the lobule-alveolar system of the udder (or mammary glands)now being complete as a result of a tirnulation produced. by thesubstance mentioned"; the next step is to initiate the milk secretionprocess in the newly grown cells of the duct and lob ulealveolar system.This is effected toy-discontinuing administration of progesterone orprogesteronelike compounds and by maintaining or increasing the amountof estrogen or estrogen-like com,- pounds. As soon as the udder fillswith mi1k', th e milk secretion process is started and continues withthe regular daily removal of -milk. 2 After about ten days the estrogenmay be discontinued or at least reduced-in amount and milk productionwill continue as it did following a normal parturition.

In normal pregnant animals theprogesterone and estrogen will be orallyadministered as described in paragraph numbered 3 under the headingGrowth of the Udder immediatelypreceding this heading. Normally, thefull growth. of the udder is developed between the first one-half ortwo-thirds period of the pregnancy. The synergistic efiect of exogenoushormones administered during this period of pregnancy having increasedthe total growth of the glandj'the mixture (feed containing estrogen andprogesterone in proper proportions) may be gradually discontinued ifpreferred, or it may be continued until 'rns'rs' WITH EXPERIMENTALAmt/mi,.s.

The growth of the mammary gland, the initiation and maintenance of themilk secretion process has been discovered to be stimulatedprogressively by the oral administration of the hormones. sinceit hasbeen observed thatmam- .mal's follow the same pattern of mammary glanddevelopment, the most complete description of our invention will bestated in terms of the requirements of the experimental animals. The

first stage of mammary development consists of the growth of the ductsystem. This type of growth can be stimulated by the following pro-'cedures.

EXAMPLE I.1MICE The duct .system of either maleor femalelmice arestimulated by incorporating into the regular complete mouse feed thedimethyl ether ofdiethylstilb-estrol in amounts varying ,from 0.0l to5.0 mg. (0.000001 to 0.000578) per kg. of feed and developed ductsystems can be produced in four to six weeks.

Other derivatives of diethylstilbestrol and related synthetic andnatural estrogenic hormones may be substituted in the above completemouse feedin amounts which will produce equal duct growth.

The synthetic estrogens depress feed consumptlon in proportion to theamounts present in the feed. Thus upon levels which produce the mostrapid duct growth as indicated above, body weight is inhibited and otherundesirable efiects are produced.

The duct system of the mouse mammary gland may also be stimulated by theincorporation in the ration of methyl testosterone in amounts varyingfrom 1.0 to 50.0 mg. per kg. (0.0001 to 0.005%) of feed and fed for fourweeks or more. Physiological equivalent amounts of other androgens orcrude extracts of cow manure may be substituted.

Furthermore, some of the undesirable effects of the diethylstilbestrolderivatives may be corrected by the simultaneous administration ofmethyl testosterone with the above synthetic estrogens. There is realsynergism of the two sex hormones approaching closely the naturalcondition. Either the estrogens alone and their derivatives or .theandrogens and their derivatives or these two sex hormones combinedtogether become more effective in stimulating duct growth when combinedwith protamone, the synthetic thyroprotein containing 3% equivalent ofthyroxine. This synergism or augmentation of the duct growth rate isaccomplished by incorporating in the complete mouse feed the amounts ofestrogens or androgens separately or together prescribed above togetherwith a range of protamone from 100 mg. to 3000 mg. per kg. of feed. Thetolerance of protamone is greater with female mice than males so thatthehigher portion of the range indicated applies only to the females. Bythe feeding of the above combinations of matter it is possible in themale, or castrate, immature or sterile female to stimulate the full ductdevelopment of the mammary gland.

EXAMPLE II.RABBITS The duct system of the immature female rabbit orimmature and mature male rabbit is stimulated to extensive growth anddevelopment by incorporating into the regular rabbit grain feed (rabbitsare normally fed grain ration and hay) the estrogen,'androgen or otherselected natural or synthetic chemical or hormones. If the hay is groundand incorporated in the grain ration the amounts of the severalingredients would be changed accordingly, the synthetic estrogen calledthe dimethyl ether of diethylstilbestrol in amounts varying from 0.4 to5.4 mg. per kg. feed. Other derivatives of diethyl stilbestrol and othersynthetic or natural estrogens may be substituted in equivalentphysiological amounts. Still larger amounts of the dimethyl ether can befed for limited periods but undesirable side effects begin to appear. Atthe upper limits recommended, namely, 5.4 mg. per kg. feed, a feedingperiod of about four weeks produces good duct development. Similar ductdevelopment may be stimulated in the rabbit by the incorporation in thegrain ration of methyl testosterone in amounts varying from 1.0 to 50mg. per kg. (0.0001 to 0.005%) of feed and fed for about four weeks.These two sex hormones may be combined in the grain ration in the abovedescribed amounts with a synergism of their separate function.

Either the estrogens alone and their derivatives or the androgens andtheir derivatives or these two sex hormones combined together becomemore effective in stimulating duct growth when combined with protamone,the synthetic thyroprotein containing 3% equivalent of thyroxine. Theaugmentation of the duct growth is accomplished by incorporating in thegrain feed the amounts of estrogens or androgens separately or togetherwith a range of protamone from 0.02 to 0.0075% or 200 to mg. per kg.grain feed. The larger amount represents the upper limits of tolerancefor the period of about four weeks.

EXAIVIPLE III.GOATS The duct systems in the udders of young goats may bestimulated to growth by incorporating in the grain ration the dimethylether of diethylstilbestrol in amounts varying from 10 to 50 mg. per kg.feed. This grain feed should be fed at the rate of about 1 kilo feed perlbs. body weight daily. Other estrogens may be substituted in equivalentphysiological amounts orally administered. This feeding program may becontinued in half grown kids for a period of two to three months.

Methyl testosterone in amounts varying from 10 to 50 mg. per kg. of feedmay be substituted for the estrogens or the two may be combined asdescribed for the mouse and rabbit. Estrogen and androgen alone ortogether may be combined within the dosage range prescribed withprotamone, the synthetic thyroprotein containing 3% equivalent ofthyroxine. The protamone is added to the grain ration at the rate of 0.5to 1.75 gm. per kg. grain mixture to be fed at the rate of 1 kg. feedper 100 lbs. of body weight per day. By this combination of hormones themaximum synergism effective in rapid gland development occurs.

EXANIPLE IV .-CATTLE In the stimulation of the duct system of heifers.the normal plan would be to wait until sexual maturity or until theanimal is a year or more of age. Even if it were impossible to causesuch animals to conceive, the duct system in each quarter would bepartially stimulated by the recurring estrus cycles. To complete thegrowth of the duct system, there should be incorporated in the grainration dimethyl ether of diethylstilbestrol in amounts ranging from 10to 200 mg. per kg. of feed. This grain is fed at the rate of 3 kg. per1000 lbs. body weight daily.

Methyl testosterone in amounts varying from 10 to 100 mg. per kg. offeed may be substituted for the estrogen or preferably the estrogenprescribed above may be combined with androgen and the combination ofmatter fed at the rate. of 3 kg. per 1000 lbs. body weight daily. Topromote maximum rapidity of gland growth, the estrogen or androgenseparately or together are combined with protamone, the syntheticthyroprotein containing 3% equivalent of thyroxine. The protamone isadded to the above described hormones and to the grain ration at therate of 1.0 gm. to 6.0 gm. per kg. of feed to be fed at the rate of 3kg. per 1000' lbs. body weight daily.

but less efficient use of the hormone. of progesterone above the levelspecified are ef-j fective but may induce degrees of anesthesia In placeof LobuZe-alveolar growth The duct system of the mammary glandsv orudders can be developed by the composition of matter described in theprevious examples. This development must take place before thelobulealvolar system can be grown. The hormone combinations previouslydescribed stimulate mammary gland developmentefliciently but willnotcarry the mammary gland development much beyond this initial stage.The compositions of matter to be described will also stimulate ductdevelopment when it is not present but will, in addition, carry themammary gland (udder) development and growth to the point comparable tothat at the period of pregnancy. The same treatment can be administeredto pregnantanimals to cause mammary gland growth in excess of their owncapacity to produce if their inheritance for gland or udder size islimited.

EXAMPLE V.' MICE The lobule-alveolar system of the mouse mammary glandis stimulated to grow from the,

duct system by the continued feeding of the dimethylether ofdiethylstilbestrol indicated above plus 0.1 to 1.0 gm. of. progesteroneper kg. of feed. Amounts of progesterone slightly above the 1.0

.gm. level may be safely fed with good results but the degree oflobule-alveolar development will not be greatly improved. progesteroneis known to possess anesthetic properties.

Thus a typical mouse ration to stimulate 'lobule-alveolar developmentwould consist of the following composition:

Dimethyl ether of diethylstilbesterol mg. per kg 0.05-1.5 Progesteronegm. per kg 0.1-1.0

Maximum synergism of the diethylstilbestrol is obtained when not overthe'upper limits of the compound indicated above are fed, i. e., 1.0 mg.per kg. of feed. If the dimethylstilbestrol' is en'- tirely eliminatedfrom the ration, progesterone alone will stimulate the lobule-alveolarsystem but the requirements are increased from to and derivatives inequal physiological amounts.

The rate and completeness of lobule-alveolar growth can be stimulated byadding to the above ration, protamone, the synthetic thyroproteincontaining 3% equivalent of thyroxine, in

amounts ranging from 100 mg. to 3000 mg. per I kg. of the abovecomposition of matter.-

EXAMPLE VI.RABBITS Growth of the lobule-alveolar system is stimulated toa degree comparable to pseudo-preg- An excess of 10 The rate andcompleteness of lobule-alveolar growth can be augmented by the additionto the ration of protamone the synthetic thyroprotein containing 3%equivalent of thyroxine, in amounts ranging from 7 5 mg. to 200 mg. perkg. of grain mixture.

EXAMPLE VII.-GOATS Growth of the lobule-alveolar system ofth'e goatsudder to a degree comparable to that of 73, of pregnancy is stimulatedby incorporating nancy .or 20 days of pregnancy by the continued feedingof the above amounts of dimethyl ether of diethylstilbestrol (0.4 to 5.4mg. per kg. of

feed). plus.(10 mg. to 300 mg. of progesterone The growth on the lowerlevels per kg. of feed).

of progesterone specified will be very slow whereas the higher. levelswill promote faster growth Amounts rivatives of the above. may besubstituted on an equal physiological basis.

in the grain ration the followingingredients:

Dimethyl ether of diethylstilbestrol at a rate of 5 mg. to 10 mg. perkg. of feed.

Progesterone at the rate of 10 mg. to 500 mg. per kg.

The grain ration is fed at the rate of about 1. kg. per lbs. body weightof the goats per day. In place of progesterone, pregneninolone,desoxycorticosterone and other related compounds and derivatives may besubstituted on an equal physiological basis. Excessive amounts of thesecompounds likewise have atendency to induce varying degrees ofanesthesia.

The rate and completeness of lobule-alveolar growth can be augmented bythe addition to the above composition of matter, protamone, thesynthetic thyroprotein containing 3% of thyroxine, in amountsrangingfrom 0.5 to 1.75 gm. per kg. grain mixture to be fed in theamounts prescribed above.

EXAMPLE VIII.-CATTLE Growth of the lobule-alveolar system ofuthecows"'udderto a degree comparable to that of pregnancy-can be stimulatedby incorporating in the grain ration the following ingredients;

Dimethyl ether of diethylstilbestrol at theirate of 10 mg. to 200 mg.per kg. of. feed. Progesterone at the rate of 10 mg. to 200 mg.

per kg. of feed.

This composition of matter is to be fed at the rate of 3 kg. per 1000lbs. body weight daily. In place of progesterone, pregneninolone,,desoxycorticosterone and other related compounds and derivatives may besubstituted on an equal physiological basis. The upper amounts .ofprogesterone prescribed may be exceeded but the ratio of estrogen toprogesterone reaches a point of maximum synergism with less rapid growthoccurring on either side. The anesthetic effect of high dosages of thesecompoundsmust be avoided.

To promote more rapid growth at the point of maximum synergism ofestrogen and progesterone it is necessary to add a third hormone,synthetic thyroprotin or its equivalent containing 3% of thyroxine.'Protamone or equivalent is added to the above described hormones and tothe grain ration at the rate of 1.0 gm. to 5.0 gm. per kg. of feed andfed at the-rate of 3 kg. per 1000 lbs. body weight daily. The feedconsumption is also favorably influenced by protamone.

The time required to cause the completedevelopment of thelobule-alveolar system will vary with the amount andsynergism of theseveral hormones prescribed. Since this development during pregnancyrequiresabout 6 months in Seattle, 2. period of similar length or longerwould pregnant animals, the change to be described will usually occurshortly before the normal period of parturition.

EXAMPLE IX.MICE

When complete lobule-alveolar mammary gland growth has been induced bythe above described treatment for periods of not less than two to threeweeks (indefinitely longer treatment may be practiced), lactation canthen be induced by the removal of the progesterone or related compoundsfrom the ration but by the continuation of the dimethyl ether ofdiethylstilbestrol for a period of two days to a week, then allestrogens are removed from the ration and lactation will occurspontaneously. Baby mice placed with such lactating mice will obtainmilk 'sufiicient to grow normally.

EXAMPLE X.RABBITS Following full lobule-alveolar growth of the mammaryglands, lactation is produced by eliminating the progesterone butcontinuing the dimethylether of diethylstilbestrol in the ration at thelevel indicated above (0.4 to 5.4 mg. per kg. of feed) for a period notexceeding a week, then all hormone treatment is discontinued and milksecretion quickly'fills the mammary glands.

EXAMPLE fir-GOATS Milk secretion is initiated in the goat by feeding agrain ratio containing the dimethyl ether of diethylstilbestrol or otherestrogens in equivalent amounts varying from to 50 mg. per kg. of feedwith protamone at the rate of 0.5 to 1.75 gm. per kg. of grain mixturefed at the rate of 1 kg. feed per 100 lbs. body weight per day for aperiod not to exceed one week. At that time the dimethyl. ether ofdiethylstilbestrol is re- EXAMPLEv XII.-CATTLE In sterileycows whosemammary glands have been developed by the methods previously describedor in pregnant cows in which the normal growth of the udder has beenaugmented by the feeding of these compositions, the secretion of milkand the parturition of pregnant cows, followed by lactation, isinitiated by removal of the progesterone from the composition used tostimulate the growth of the lobule-alveol'ar sys- 12 tem. Thecomposition fed would then be composed as follows:

Dimethyl ether of diethylstilbestrol (or equivalent) at the rate of 10mg. to 200 mg. per kg. of feed and fed at the rate of 3 kg. per 1000lbs; body Weight daily.

In pregnant cows this composition is fed until after parturition.Sterile cows would be fed for a week or 10 days while regular milkingwould be started. When milk production reaches a peak, the maintenanceof this level of milk secretion is aided by the followingadministrations:

Dimethyl ether of diethylstilbestrol, 2.5 mg. to

50.0 mg. per kg. of feed Protamone (or equivalent), 1.0 gm. to 5.0 gm.

per kg. of feed Methyl testosterone (or equivalent), 5.0 mg.

50 mg. per kg. of feed These to be fed at the rate of 3 kg. per 1000lbs. body weight daily so long as profitable milk production ismaintained. In sterile cows, the growth of the lobule-alveolar systemcan then be reinitiated by the administration of the compositionpreviously described which includes progesterone.

In lactating cows an alternative method of stimulating the growth of thelobule-alveolar system of the udder could be to substitute thecomposition described under lobule-alveolar growth for periods of onemonth or more to rejuvenate the gland, then return to the lactationstimulating ration.

As pointed out hereinbefore, there may be used in lieu of progesteroneany of the various compounds having the property of producingprogestational proliferation of the rabbit uterus, and in the appendedclaims the phrase compound having progestational producing activity ismeant to embrace all such compounds, but no others.

Having thus described my invention, I claim:

1. A method for stimulating the complete growth of the mammary glands inmilk secreting animals, comprising the stepsof adding to their normalfeed ration during pregnancy and prior to parturition, a mixture ofsynergistic compounds including estrogens and progesterone.

2. A method as in claim 1 including the step of discontinuing .feedingof the progesterone when development of the mammary glands is completeand increasing the amount oi estrogen until milk secretion proceedsnormally.

3.' A .method as in claim 1 wherein pregneninolone is substituted forthe progesterone.

4. A method for stimulating the complete growth of the mammary glandsand in milk secreting animals, comprising the step of adding to theirnormal food ration during pregnancy and .prior to parturition,synergistic compounds including an estrogen anda compound havingprogestational producing activity.

5. A method for favorably affecting the physiological processes .ofmammals, comprising the steps. of adding to, their normal feed ration acombination of synergistic compounds including 6. A method for favorablyafiecting the physiological processes of lactating animals comprisingthe steps of adding to their normal feed ration a combination ofsynergistic compounds including an estrogen and a compound havingprogestational producing activity thereby to stimulate the completegrowth of the mammary gland, discontinuing the compound havingprogestational producing activity when the growth of the mammary glandis complete, and thereafter adding to the normal feed ration acombination of synergistic compounds comprising an estrogen and anandrogen.

7. A method for favorably afiecting the physiological processes oflactating animals comprising the steps of adding to the normal feedration a combination of synergistic compounds including an estrogen anda compound having progestational producing activity thereby to stimulatethe complete growth of the mammary gland, discontinuing the compoundhaving progestational producing activity when the growth of the mammarygland is complete, and thereafter adding to the normal feed ration acombination of synergistic compounds comprising an estrogen and anandrogen, and a thyroxine containing material.

8. A method for favorably affecting the physiological processes ofpregnant lactating animals comprising the steps of adding to theirnormal feed ration a combination of synergistic compounds including anestrogen and a compound having progestational producing activity therebyto stimulate the complete growth of the mammary glands, discontinuingthe compound having progestational producing activity prior toparturition and thereafter adding an estrogen to the normal feed ration.

9. A method for favorably afiecting the physiological processes ofpregnant lactating animals comprising the steps of adding to theirnormal feed ration a combination of synergistic compounds including anestrogen and a compound having progestational producing activity therebyto stimulate the complete growth of the mammary glands, discontinuingthe compound having progestational producing activity prior toparturition and thereafter adding to the normal feed ration acombination of synergistic compounds including an estrogen and anandrogen.

10. A method for favorably affecting the physiological processes ofpregnant lactating animals comprising the steps of adding to theirnormal feed ration a combination of synergistic compounds including anestrogen and a compound having progestational producing activity therebyto stimulate the complete growth of the mammary glands, discontinuingthe compound having progestational producing activity prior toparturition and thereafter adding to the normal feed ration acombination of synergistic compounds including an estrogen, an androgenand a thyroXine-containing compound.

11. A method for artificially stimulating the complete cycle of mammarygland development and lactation of mammals as observed during recurringestrous cycles, comprising the consecutive steps of first adding totheir normal feed ration a combination of synergistic compoundsincluding an estrogen and an androgen thereby to develop the mammaryduct system, then adding to their normal feed ration a compound havingprogestational producing activity until growth of the mammary glands iscomplete, and then discontinuing the addition of the last mentionedcompound and adding to the normal feed ration a thyroXine-containingmaterial to' augment the milk secretion.

CHARLES W. TURNER.

REFERENCES CITED The following references are of record in the file ofthis patent:

Chem. Ab., vol. 36, 1942, page 3832 Chem. Ab., vol. 41, 1947, page3854*.

Chem. Ab., vol. 38, page 1014*.

Chem. Ab., vol. 38, page 407".

Chem. Ab., vol. 40, page 6601 Chem. Ab., v01. 33, page 8759 Chem. Ab.,vol. 31, page 6303 Chem. Ab., vol. 40, 1946, page 1573.

Chem. Ab., vol. 40, 1946, page 4126*.

Chem. Ab., vol. 38, 1944, page 5278 Chem. Ab., Vol. 37, 1943, page3491".

Annual Review of Biochemistry, vol. XIV, 1945, pages 578 and 579.

Artificial Induction of Lactation in Bovines, by Folley et al., pages 24to 36, Journal of Endocrinology, vol. 4, 1944.

1. A METHOD FOR STIMULATING THE COMPLETE GROWTH OF THE MAMMARY GLANDS INMILK SECRETING ANIMALS, COMPRISING THE STEPS OF ADDING TO THEIR NORMALFEED RATION DURING PREGNANCY AND PRIOR TO PARTURITION, A MIXTURE OFSYNERGISTIC COMPOUNDS INCLUDING ESTROGENS AND PROGESTERONE.